We have found that a group of new substituted azetidinones are potent elastase inhibitors and therefore are useful anti-inflammatory and antidegenerative agents.
Proteases from granulocytes and macrophages have been reported to be responsible for the chronic tissue destruction mechanisms associated with inflammation, including rheumatoid arthritis and emphysema. Accordingly, specific and selective inhibitors of these proteases are candidates for potent anti-inflammatory agents useful in the treatment of inflammatory conditions resulting in connective tissue destruction, e.g. rheumatoid arthritis, emphysema, bronchial inflammation, chronic bronchitis, glomerulonephritis, osteoarthritis, spondylitis, lupus, psoriasis, atherosclerosis, sepsis, septicemia, shock, myocardial infarction, reperfusion injury, periodontitis, cystic fibrosis and acute respiratory distress syndrome.
The role of proteases from granulocytes, leukocytes or macrophages are related to a rapid series of events which occurs during the progression of an inflammatory condition:
(1) There is a rapid production of prostaglandins (PG) and related compounds synthesized from arachidonic acid. This PG synthesis has been shown to be inhibited by aspirin-related nonsteroidal anti-inflammatory agents including indomethacin and phenylbutazone. There is some evidence that protease inhibitors prevent PG production; PA1 (2) There is also a change in vascular permeability which causes a leakage of fluid into the inflamed site and the resulting edema is generally used as a marker for measuring the degree of inflammation. This process has been found to be induced by the proteolytic or peptide cleaving activity of proteases, especially those contained in the granulocyte, and thereby can be inhibited by various synthetic protease inhibitors, for example, N-acyl benzisothiazolones and the respective 1,1-dioxides. Morris Zimmerman et al., J. Biol. Chem., 255, 9848 (1980); and PA1 (3) There is an appearance and/or presence of lymphoid cells, especially macrophages and polymorphonuclear leukocytes (PMN). It has been known that a variety of proteases are released from the macrophages and PMN, further indicating that the proteases do play an important role in inflammation. PA1 (1) Recent histochemical investigations showed the accumulation of granulocytes at the cartilage/pannus junction in rheumatoid arthritis; and PA1 (2) a recent investigation of mechanical behavior of cartilage in response to attack by purified elastase demonstrated the direct participation of granulocyte enzymes, especially elastase, in rheumatoid cartilage destruction. H. Menninger etal., in Biological Functions of Proteinases, H. Holzer and H. Tschesche, eds. Springer-Verlag, Berlin, Heidelberg, New York, pp. 196-206, 1979. PA1 R.sup.1 is C.sub.1-6 alkyl or C.sub.1-6 alkoxy-C.sub.1-6 alkyl; PA1 M is PA1 Ra and Rb are each individually PA1 R.sup.2 and R.sup.3 are each independently PA1 R.sub.4 is (a) ##STR9## or (b) ##STR10## where R.sub.x is carboxy C.sub.1-6 alkyl, benzyloxycarbonylC.sub.1-3 alkyl, or t-butoxycarbonylC.sub.1-3 alkyl, PA1 Q is a covalent bond or ##STR11## wherein R.sub.5 and R.sub.6 are each individually C.sub.1-3 alkyl or,hydrogen, PA1 Y is ##STR12## or a covalent bond; R.sub.12 is hydrogen or C.sub.1-3 alkyl; PA1 R.sub.7 and R.sub.8 are each individually PA1 X.sub.1 is PA1 n is 1, 2, 3, 4 or 5; PA1 R.sub.9 is selected from hydrogen, C.sub.1-4 alkyl, and C.sub.1-3 alkoxyC.sub.1-3 alkyl; or phenyl, phenyl C.sub.1-3 alkyl, pyridyl, and pyridyl C.sub.1-3 alkyl; PA1 R.sub.10 and R.sub.11 are each independently selected from hydrogen, C.sub.1-4 alkyl, and C.sub.1-3 alkoxy C.sub.1-3 alkyl, or aryl as defined above, or are together O.dbd.; or PA1 R.sub.7 and R.sub.8 are joined together to form mono or di substituted ring of 4, 5, 6, or 7 atoms or 7 to 12 atoms such as PA1 wherein R.sub.9 and R.sub.12 are joined together to form a mono or di substituted saturated monocyclic ring of 5, 6, or 7 atoms, said ring having one hetero atom which is the nitrogen to which R.sub.9 is attached; or wherein R.sub.10 and R.sub.12 are joined together to form a mono or di substituted saturated monocyclic ring of 5, 6, or 7 carbon atoms; or PA1 wherein R.sub.8 and R.sub.11 are joined together to form a mono or di substituted saturated monocyclic ring of 5, 6, or 7 atoms, said ring having one hetero atom which is the nitrogen to which R.sub.8 is attached; and the substituents are independently selected-from Hydrogen and C1-3alkyl. PA1 R.sup.1 is C.sub.1-6 alkyl or C.sub.1-6 alkoxy-C.sub.1-6 alkyl; PA1 M is PA1 Ra is PA1 Rb is hydrogen, or C.sub.1-6 alkyl, PA1 R.sup.2 and R.sup.3 are each independently PA1 R.sub.4 is (a) ##STR17## or (b) ##STR18## where R.sub.x is carboxyC.sub.1-6 alkyl, benzyloxycarbonylC.sub.1-3 alkyl, or t-butoxycarbonylC.sub.1-3 alkyl, PA1 Q is a covalent bond or ##STR19## wherein R.sub.5 and R.sub.6 are each individually C.sub.1-3 alkyl or hydrogen PA1 Y is ##STR20## or a covalent bond; R.sub.12 is hydrogen or C.sub.1-3 alkyl; PA1 R.sub.7 and R.sub.8 are each individually PA1 X.sub.1 is PA1 X.sub.2 is hydrogen, halo or C.sub.1-6 alkyl; PA1 n is 1, 2, 3, 4 or 5; PA1 R.sub.9 is selected from hydrogen, C.sub.1-4 alkyl, and C.sub.1-3 alkoxyC.sub.1-3 alkyl; PA1 R.sub.10 nd R.sub.11 are each independently selected from hydrogen, C.sub.1-4 alkyl, and C.sub.1-3 alkoxy C.sub.1-3 alkyl; or PA1 R.sub.7 and R.sub.8 are joined together to form mono or di substituted ring of 4, 5, 6, or 7 atoms such as PA1 wherein the substituents are each selected from the group consisting of hydrogen and C.sub.1-3 alkyl, benzyloxycarbonyl, carboxy, phenyl C.sub.1-3 alkyl amino carbonyl, pyrrolidinyl, methyl, hydroxy C.sub.1-3 alkyl, C.sub.1-6 alkyloxy, C.sub.1-4 alkyloxy carbonyl, and oxo; or R.sub.8 and R.sub.9 are joined together to form a saturated ring of 5 to 7 atoms and having two hetero atoms; or R.sub.9 and R.sub.10 are joined together to form a saturated ring of 5 to 7 atoms and having one hetero atom; or wherein R.sub.9 and R.sub.12 are joined together to form a ring of 5, 6, or 7 atoms, said ring being saturated; or wherein R.sub.10 and R.sub.12 are joined together to form a ring of 5, 6, or 7 atoms, said ring being saturated; or PA1 wherein R.sub.8 and R.sub.11 are joined together to form a ring of 5, 6, or 7 atoms, said ring being saturated and having one hetero atom. PA1 R is C.sub.1-3 alkyl; PA1 R.sub.1 is C.sub.1-3 alkyl; PA1 M is PA1 R.sup.2 is PA1 R.sup.3 is hydrogen, or PA1 R.sup.2 and R.sup.3 are joined together to form a methylenedioxy group or a furan ring; R.sub.5 and R.sub.6 are each individually hydrogen or C.sub.1-3 alkyl; PA1 wherein X.sub.1 is hydrogen and X.sub.2 is PA1 n is 1, 2 or 3, and PA1 R.sub.9, R.sub.10 and R.sub.11 are each independently selected from hydrogen, C.sub.1-4 alkyl, and C.sub.1-3 alkoxy C.sub.1-3 alkyl; or PA1 R.sub.7 and R.sub.8 are joined together to form a substituted ring selected from PA1 R.sub.8 and R.sub.9 are joined together to form a ring of 6 to 7 atoms and having two hetero atoms; PA1 R.sub.9 and R.sub.10 are joined together to form a saturated ring of 5 to 7 atoms and having one hetero atom; or wherein R.sub.9 and R.sub.12 are joined together to form a ring of 5, 6, or 7 atoms, said ring being saturated; or wherein R.sub.10 and R.sub.12 are joined together to form a ring of 5, 6, or 7 atoms, said ring being saturated; or PA1 wherein R.sub.8 and R.sub.11 are joined together to form a ring of 5, 6, or 7 atoms, said ring being saturated and having one hetero atom. PA1 Q is a covalent bond; PA1 R is methyl or ethyl; PA1 R.sub.1 is methyl or ethyl; PA1 M is PA1 R.sup.2 is PA1 R.sup.3 is hydrogen, or PA1 R.sup.2 and R.sup.3 are joined together to form a furan or dioxacyclopentane ring; PA1 n is 1 or 2; PA1 R.sub.9 and R.sub.10 are each independently selected from PA1 R.sub.7 and R.sub.8 are each independently selected from PA1 R.sub.7 and R.sub.8 are joined together to form a substituted ring selected from PA1 R.sub.8 and R.sub.9 are joined together to form a piperazine ring. PA1 co-administration to a patient in need of such inhibition of a therapeutically effective amount of compound selected from the group consisting of epsilon-aminocaproic acid, heparin, trasylol, prednisolone, cytosine arabinoside, .beta.-mercaptopurine, cytarabine, an anthracycline, and a vitamin A derivative;and a therapeutically effective amount of compound of Formula I as defined above.
In general, proteases are an important family of enzymes within the peptide bond cleaving enzymes whose members are essential to a variety of normal biological activities, such as digestion, formation and dissolution of blood clots, the formation of active forms of hormones, the immune reaction to foreign cells and organisms, etc., and in pathological conditions such as the degradation of structural proteins at the articular cartilage/pannus junction in rheumatoid arthritis etc.
Elastase is one of the proteases. It is an enzyme capable of hydrolyzing the connective tissue component elastin, a property not contained by the bulk of the proteases present in mammals. It acts on a protein's nonterminal bonds which are adjacent to an aliphatic amino acid. Neutrophil elastase is of particular interest because it has the broadest spectrum of activity against natural connective tissue substrates. In particular, the elastase of the granulocyte is important because, as described above, granulocytes participate in acute inflammation and in acute exacerbation of chronic forms of inflammation which characterize many clinically important inflammatory diseases.
Proteases may be inactivated by inhibitors which block the active site of the enzyme by binding tightly thereto. Naturally occurring protease inhibitors form part of the control or defense mechanisms that are crucial to the well-being of an organism. Without these control mechanisms, the proteases would destroy any protein within reach. The naturally occurring enzyme inhibitors have been shown to have appropriate configurations which allow them to bind tightly to the enzyme. This configuration is part of the reason that inhibitors bind to the enzyme so tightly (see Stroud, "A Family of Protein-Cutting Proteins" Sci. Am. July 1974, pp. 74-88). For example, one of the natural inhibitors, .alpha..sub.1 -Antitrypsin, is a glycoprotein contained in human serum that has a wide inhibitory spectrum covering, among other enzymes, elastase both from the pancreas and the PMN. This inhibitor is hydrolyzed by the proteases to form a stable acyl enzyme in which the active site is no longer available. Marked reduction in serum .alpha..sub.1 -antitrypsin, either genetic or due to oxidants, has been associated with pulmonary emphysema which is a disease characterized by a progressive loss of lung elasticity and resulting respiratory difficulty. It has been reported that this loss of lung elasticity is caused by the progressive, uncontrolled proteolysis or destruction of the structure of lung tissue by proteases such as elastase released from leukocytes. J. C. Powers, TIBS, 211 (1976).
Rheumatoid arthritis is characterized by a progressive destruction of articular cartilage both on the free surface bordering the joint space and at the erosion front built up by synovial tissue toward the cartilage. This destruction process, in turn, is attributed to the protein-cutting enzyme elastase which is a neutral protease present in human granulocytes. This conclusion has been supported by the following observations:
In a second aspect this invention concerns the use of novel azetidinones in the treatment of certain cancers including nonlymphoblastic leukemias, acute myelogenous leukemia (FAB M1 and FAB M2), acute promyelocytic leukemia (FAB M3), acute myelomonocytic leukemia (FAB M4), acute monocytic leukemia (FAB M5), erythroleukemia, chronic myelogenous leukemia, chronic myelomonocytic leukemia, chronic monocytic leukemia and conditions associated with leukemia involving activity of PMN neutral proteases e.g. disseminated intravascular coagulation. We have found that the substituted azetidinones disclosed herein are inhibitors of proteinase 3 (PR-3), also known as myeloblastin.
See C. Labbaye, et al., Proc. Natl. Acad. Sci. USA, vol. 88, 9253-9256, (1991), Wegner autoantigen and myeloblastin are encoded by a single mRNA; D. Campanelli, et al., J. Exp. Med., vol. 172, 1709-1714, (1990), Cloning of cDNA for proteinase 3: A serine protease, antibiotic, and autoantigen from human neutrophils; and Bories, st. al., Cell vol. 59, 959-968, (1989) Down-regulation of a serine protease, myeloblastin, causes growth arrest and differentiation of promyelocytic leukemia cells.
Recently, down regulation of PR-3 has been implicated in the proliferation and maintenance of a differentiated state of certain leukemia cells. In particular, Bories, et. al., have shown that expression of this enzyme, hereinafter designated proteinase 3/myeloblastin, can be inhibited by treatment of HL-60 human leukemia cells with an antisense oligodeoxynucleotide and that such treatment induces differentiation and inhibits proliferation of these cells. Moreover, we have now demonstrated that the treatment of the HL-60 cell human leukemia cell line, among others, with the compounds of the instant invention, likewise results in the inhibition of proliferation and induction of differentiation in such cells.
Accordingly, we believe that treatment of leukemia such as nonlymphoblastic leukemias, acute myelogenous leukemia (FAB M1 and FAB M2), acute promyelocytic leukemia (FAB M3), acute myelomonocytic leukemia (FAB M4), acute monocytic leukemia (FAB M5), erythroleukemia, chronic myelogenous leukemia, chronic myelomonocytic leukemia, chronic monocytic leukemia and conditions associated with leukemia involving activity of PMN neutral proteases e.g. disseminated intravascular coagulation, comprising: administration of a therapeutically effective amount of compound of Formula I will result in remission of the disease state. Administration may be either oral or parenteral.